Therapeutic composition and process of making it



Patented Na -1, i939 'umrso STATE s PATEN'T orrics' 2,179,384 THERAPEUTIC COMPOSITION AND PROCESS OF MAKIN G IT No Drawing. Application April 5, 1931, Serial No. 135,106. Renewed April 18, 1930 v 10 Qlaims.

it is the object oi our invention to enhance utility of Insulin-protamine compositions.

It has been denmonstrated by Hagedorn and has co-workers that the blood-sugar-lowering action of Insulin" may be prolonged by using protamine with the Insulin. See The Journal of the American Medical Association for January 18, 1036, volume 106, page 1'77.

Experience has shown that all or a part of Insulin"-containing precipitates in lnsulin"- protarnine compositions will often adhere to the inside of containers in which the compositions are stored. Experience, has shown, also, that in lnsulin-protamine compositions the precipitates commonly form aggregates which cannot readily be adequately disintegrated. The utility of Insulin-protamine compositions in which precipitates either form such aggregates or adhere to the inside of containers is limited, since obviously the precipitates in the compositions should remain completely and homogeneously suspended during withdrawal of doses of the compositions.

We have discovered that such limitation oi utility of Insu1in-protamine compositions can be avoided, and in consequence that utility of such compositions can be enhanced, by having present in the compositions an adequate amount of metal of the class consisting of zinc, cadmium, nickel, and cobalt. Also, we have discovered that by the presence of this amount of metal of the class defined the period of utility of Insulin"- protamine compositions is greatly lengthened. Moreover, we find that by the presence of this amount of either zinc or cadmium in Insulinprotamine compositions the prolonged bloodsugar-lowering action of the compositions is eittended. We here assert no theory as to the nature of the actions of the metal present, or as to whether or not there is any chemical combination between the metal and either or both of the Insulin and the protamine components.

According to our invention, in preparing Insulin-protamine compositions we ensure the presence in the compositions of an amount of zinc, cadmium, nickel, and/or cobalt, per unit of Insulin", that is larger than the amount of such metal present per' unit of Insulin" in ordinary preparations of Insulin or in Insulin-protamine compositions as made prior to our discovery,

and that is at least twice as large, molecularly, as

the amount of such metal present per unit of Insulln" in crystals of "Insulln. ["Insulin crystallized with zinc contains about 0.52% of zinc, Insulin" crystallized. with cadmium contains about 0.77% of cadmium, Insulin crystallized with nickel contains about 0.41% of nickel, and Insulin crystallized with cobalt contains about 0.44% of cobalt, in terms or dry weight of the crystals] Beneficial results are obtainable by. having present in Insulin-protamine compositions as little as half a milligram of zinc (or molecularly equivalent amounts if the other metals are used) per thousandunits oi the Insulin; and desirably, in order to ensure that not less than this quantity of metal be present, we add at least this quantity. We may add any larger quantity of metal provided only that the quantity added be not so large as to induce untoward reactions, such as undue pain from injections. We have obtained satisfactory results from adding 20 me. of zinc (and also'from adding molecularly equivalent quantities of the other metals) per 1000 units of the insulin. Most desirably we ensure the presence oi a total of about 2 mg. of zinc per 1000 units oi the lnsu1in".

Illustrations oi our invention are as follows:

In any desired way we make a preparation of Insulin and protamine-as by adding to a solution of Insulin any suitable protamine, such as salmiridine or scombrine or s'almine, commonly in the range of about 2 to do mg. of protamine per 1000 units of the llnsu1in". During or following the operations of malrlng this preparation, we add zinc in an amount rinsing between about i and about 5 ma. per 1000 units of the Insulin. Instead oi zinc, we may add cadmium; and instead oi either zinc or cadmium, we may add nickel or cobalt; but zinc is our first choice, and cadmium our second. Instead of only one of these four metals, we may add two or more. Preferably we add metal of the defined class in the form of chlorides, but we may addmetal in elemental form, or in other forms, such as ace-r tates, oxides or hydroxides. Further, it is possi-.- ble to ensure the presence of suficient metal by conducting some of the production operations in equipment made of or containing the desired. metal or metals.

In preparing our new compositions the hydrogen-ion concentrations of the several components and of their mixtures are not of prime importance, but in making such compositions available ior clinical use and in using these, we find it desirable to attain and maintain a reaction which is on the alkaline side of the isoelectric point of the Insulin, desirably a reaction of between pH 6.9 and pHFUi. The desired reaction may be attained by adding an appropriate solution, such as a solution of a suitable buffer (a lid phosphate bufler for example), to one or more of the components or to our otherwise completepreparation.

We claim as our invention:

1. The process 0! producing an "Insulin"- protamine composition, which consists in putting Insulin and protamine together, and ensuring the presence therewith of an amount of metal of the class consisting of zinc, cadmium, nickel, and cobalt sumcient to correspond molecular?! to at least half a milligram of zinc per thousand units of the Insulin.

2. The process of producing an Insulin"- protamine composition, which consists in adding a protamine to Insulin, and also adding metal of the class consisting of zinc, cadmium, nickel, and cobalt.

3. The process of producing an Insuiin"- protamine composition, which consists in adding a protamine to Insulin", and also adding metal of the class consisting of zinc, cadmium, nickel, and cobalt in suflicient amount to make the total quantity of such metal present per unit of Insulin" at least twice as large molecularly as the quantity of metal present per unit of Insulin'' in crystals of Insulin.

4. The process of producing an Insulin"- protamine composition, which consists in adding to Insulin" a protamine and zinc.

5. The process of producing an Insulin protamine composition, which consists in adding to Insulin" a protamine and cadmium.

6. A composition of matter, comprising "Insulin", protamine, and added metal of the. class consisting of zinc, cadmium, nickel, and cobalt.

7. A composition of matter, comprising Insulin, protamine, and an amount of metal of the class consisting of zinc, cadmium, nickel, and cobaltsumcient to correspond molecularly to at least half a milligram of zinc per thousand units of the Insulin.

8. A composition of matter, comprising "Insulinf, protamine, and a quantity of metal of the class consisting of-zinc, cadmium, nickel, and cobalt which per unit of Insulin is at least twice as large molecularly as the quantity of metal present per unit of Insulin" in crystals of Insulin".

9. Acomposition of matter, comprising "Insulin", protamine, and added zinc.

10. A composition of matter, comprising Insulin, protamine, and added cadmium.

DAVID ALYMER SCOTT. ALBERT MADDEN FISHER. 

